Hi! I'm trying to process .fasta.gz files to .bam files, and then I'm uploading them into SpliceWiz in order to do alternative splicing analysis. Every time I get to the stage where I filter the AS events in SpliceWiz, the system crashes. I've pasted the code that it outputs as it crashes here. To make the .bam files, I am uploading SRA accession numbers to Galaxy, trimming using CutAdapt, and then using RNA STAR to generate the .bam file. The files upload into SpliceWiz without any problem, but they have zero values in most of the columns when I review the .bam file (in directionality_strength, intergenic_fraction, etc.). Would anyone be able to help me figure out which part of this process I'm doing wrong, or could someone suggest a better method to perform alternative splicing analysis? I have limited experience coding, but I'm not a complete novice.

Code should be placed in three backticks as shown below

Listening on http://127.0.0.1:5981
Feb 27 13:20:32 Loading NxtSE object from file...
Feb 27 13:20:33 ...loading reference (NxtSE)
Feb 27 13:20:41 ...linking assays
Feb 27 13:20:41 ...linking COV files
Feb 27 13:20:41 ...loading rowData
Feb 27 13:20:49 ...realizing NxtSE object
Feb 27 13:20:49 ...removing overlapping introns...
Feb 27 13:20:50 NxtSE loaded
Populating drop-down box with 78932 genes
Refreshing filters
Filtered 521284 ASE events
Populating drop-down box with 78932 genes
Warning: Error in prcomp.default: cannot rescale a constant/zero column to unit variance
  108: stop
  107: prcomp.default
  105: QC_PCA
  103: renderPlotly
  102: func
   99: shinyRenderWidget
   98: func
   85: renderFunc
   84: output$qc-QC_plot
    3: shiny::runApp
    2: runGadget
    1: spliceWiz
Refreshing filters
Running Depth filter
Running Participation filter
Running Participation filter
Running Consistency filter
Running Terminus filter
Running ExclusiveMXE filter
Running StrictAltSS filter
Running Protein_Coding filter
Filtered 0 ASE events
Populating drop-down box with 78932 genes
Warning: Error in [[: subscript out of bounds
  3: shiny::runApp
  2: runGadget
  1: spliceWiz
Error: invoke_wrapped: throwing std::runtime_error
> 

sessionInfo( )